Ubiquitin variant modulators of hect e3 ligases and their uses

Abstract

The invention provides ubiquitin variants that specifically bind to HECT E3 ligases, and methods of using these variants to modulate the activity of HECT E3 ligases.

Description

FIELD OF THE INVENTION[0001]This invention relates to ubiquitin variants that specifically bind to HECT E3 ligases, and use of these variants to modulate the activity of HECT E3 ligases.BACKGROUND OF THE INVENTION[0002]Ubiquitination mediated by E1-E2-E3 multi-enzyme cascades rivals phosphorylation as a predominant mechanism regulating myriad protein functions (Cohen and Tcherpakov, 2010; Nalepa et al., 2006). Repeated catalytic cycles result in substrates modified on multiple lysines with various polyubiquitin chains, which alter protein functions in an extraordinary variety of ways. Because E3 ligases control substrate specificity and the topology of ubiquitination, they represent attractive targets for therapeutic intervention (Nalepa et al., 2006; Petroski, 2008). Yet, identifying the diversity of mechanisms regulating E3 ligases, as well as generation of tools for their manipulation, has lagged behind deciphering regulation and developing therapeutics for kinases (Cohen and Tch...

AI Technical Summary

Benefits of technology

The invention provides ubiquitin variant (Ubv) polypeptides that have been modified in certain regions of the ubiquitin molecule. These modifications can include the addition or replacement of certain amino acids. The Ubv polypeptides can be used for various purposes, such as research and development, and can have improved stability and function compared to native ubiquitin. The technical effect of the invention is the creation of novel Ubv polypeptides with improved properties that can be used in various applications.

Problems solved by technology

Development of agents to selectively modulate HECT E3s has been hampered by extreme inter-domain rotations accompanying catalysis, a shallow active site, and dynamic regulation of HECT E3 activity (Escobedo et al., 2014; Gallagher et al., 2006; Huang et al., 1999; Kamadurai et al., 2013; Kamadurai et al., 2009; Mari et al., 2014; Persaud et al., 2014; Ronchi et al., 2013; Verdecia et al., 2003; Wiesner et al., 2007).

However, existing molecules generally do not conform to a general strategy that could be used to interrogate HECT E3s across the family, fall short in terms of potency and specificity, and generally have had limited utility in probing unknown HECT mechanisms.

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